Analyses Of Expression Of Human Acetaldehyde Dehydrogenase2and Function In The Mitigation Of Mice With Acute Alcoholism

Human mitochondrial aldehyde dehydrogenase2(ALDH2) is one of the keyenzymes in the human metabolic pathway of ethanol. Acetaldehyde translatedfrom ethanol is decomposed by ALDH, which reduce the harm to the humanand protect the body. Other aspects of the effect of ALDH on homan body isstudied now.There are two parts in this study: cloning and expressing of humanALDH2and effect of ALDH2which was immobilized on drunk mice.The main results were as follow:1． P. pastoris SMD1168(pPIC9K-ALDH2) strains were constructedsuccessfully through several steps. Firstly, ALDH2plasmid (pPIC9K-ALDH2)was constructed by means of genetic engineering. Secondly, ALDH2plasmid(pPIC9K-ALDH2)was switched into Pichia pastoris SMD1168byelectroporation.2．The most suitable condition of the induction process of the fermentationwere as follows: the pH of the medium was7.0, the initial induction OD600was12, cultivation temperature was28℃, the complement of methanolconcentration was1.5%(v/v) in every24hours. The activity of human ALDH2was0.122U/ml.3. supernant of the fermentation which was collected after72hoursinduction fermentation was separated and purified by Ni2+-affinitychromatography and then ALDH2protein was acquired. To get theimmobilized powder, ALDH protein freezed and dried.4. With immobilized enzyme powder and extract of Natural Products,aspartate aminotransferase and alanine aminotransferase content in boold andliver was studied. Druck and sober time of mice was also tested. It showed thatsober time of mice was shortened and aminotransferase content was decreased.